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Molecular characterization of a family of metalloendopeptidases from the intestinal brush border of Haemonchus contortus

Published online by Cambridge University Press:  02 June 2006

G. F. J. NEWLANDS ,
P. J. SKUCE ,
A. J. NISBET ,
D. L. REDMOND ,
S. K. SMITH ,
D. PETTIT  and
W. D. SMITH
G. F. J. NEWLANDS
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 OPZ, Scotland
P. J. SKUCE
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 OPZ, Scotland
A. J. NISBET
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 OPZ, Scotland
D. L. REDMOND
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 OPZ, Scotland
S. K. SMITH
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 OPZ, Scotland
D. PETTIT
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 OPZ, Scotland
W. D. SMITH
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 OPZ, Scotland
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Abstract

Substantial protection against the economically important parasitic nematode Haemonchus contortus has been achieved by immunizing sheep with a glycoprotein fraction isolated from the intestinal membranes of the worm (H-gal-GP). Previous studies showed that one of the major components of H-gal-GP is a family of at least 4 zinc metalloendopeptidases, designated MEPs 1–4. This paper describes aspects of the molecular architecture of this protease family, including the proteomic analysis of the MEP fraction of the H-gal-GP complex. These enzymes belong to the M13 zinc metalloendopeptidase family (EC 3.4.24.11), also known as neutral endopeptidases or neprilysins. The sequences of MEPs 1 and 3 suggested a typical Type II integral membrane protein structure, whilst MEPs 2 and 4 had putative cleavable signal peptides, typical of secreted proteins. Proteomic analysis of H-gal-GP indicated that the extracellular domain of all 4 MEPs had been cleaved close to the transmembrane region/signal peptide with additional cleavage sites mid-way along the polypeptide. MEP3 was present as a homo-dimer in H-gal-GP, whereas MEP1 or MEP2 formed hetero-dimers with MEP4. It was found that expression of MEP3 was confined to developing 4th-stage larvae and to adult worms, the stages of Haemonchus which feed on blood. MEP-like activity was detected in the H-gal-GP complex over a broad pH range (5–9). Since all 4 MEPs must share a similar microenvironment in the complex, this suggests that each might have a different substrate specificity.

Keywords

Haemonchus contortusglycoprotein complexproteasemetallopeptidase
Type
Research Article
Information
Parasitology , Volume 133 , Issue 3 , September 2006 , pp. 357 - 368
Copyright
2006 Cambridge University Press

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